Objectives:The aim of the present study was to use pulsed-field gel electrophoresis (PFGE) to characterise glycopeptide resistant E. faecium (GREfm) and glycopeptide resistant E. faecalis (GREfs) isolates from clinical samples. The van genotypes of the GRE isolates and the virulence factor genes, gel E, hyl, asa1, esp and cyl A were detected by multiplex PCR, hybridisation and sequenced.

Methods and Results: A total of 138 GRE recovered from 127 patients were collected between April 2004 and January 2009 from five Picardie area hospitals (France). The patients were 67 (52.7%) men and 60 (47.3%) women. The distribution of patients according to the positive samples for GRE showed that, 94 patients had one rectal swab each positive, and 1 patient presented 3 positive rectal swabs; 24 patients had one positive clinical sample each, and 8 patients had 11 positive clinical samples + 6 rectal swabs. The molecular identification showed that, 131 enterococci isolates belonged to the species E. faecium (94.9%) and 7 (5.1%) isolates to the species E. faecalis. All these GRE isolates had only the van A gene. Multiplex PCR showed that, the genotype efm A+ hyl+ esp+ and the genotype efm A+ hyl+ were found in 89, 67.9% and 42, 32.1% of the 131 clinical strains GREfmA respectively, where there were: rectal swabs, 76.4% (100/131) and clinical samples, 23.6% (31/131) of whom: deep pus, urine, blood, bile, sputum, catheter, and urethral swab. The efs A gene is frequently associated with gel E or/and asa1 genes while the efmA gene is frequently associated with hyl or/and esp. The cyl A gene was not detected in this study (Table 1). PFGE revealed seven different pulsotypes, designated A to G. Pulsotype A included 131 GREfm A genetically indistinguishable. The pulsotypes B, C, D, E, F and G included each one GREfs A isolate, which were considered as unrelated.

Conclusion: Our results show high GRE fecal carriage rate, Efm A being the mainly encountered GRE. An emerging Efm A clone has been identified during hospital Picardie area outbreaks. In contrast, only 7 Efs A have been isolated in our institutions. Molecular analysis showed the intra-hospital spread of gel E-, asa1-, hyl-, esp-positive GRE clones.